Production of modified polysaccharide S-7

a technology of polysaccharide and polysaccharide, which is applied in the field of polysaccharide s7, can solve the problems of not knowing the basis of the change in physical properties

Inactive Publication Date: 2004-03-23
SHIN ETSU BIO INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At this time we do not know the chemical structure of the derived polysacc

Method used

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  • Production of modified polysaccharide S-7
  • Production of modified polysaccharide S-7
  • Production of modified polysaccharide S-7

Examples

Experimental program
Comparison scheme
Effect test

example 2 (

GENETIC MODIFICATIONS)

Preparation ofa library of S7 genes. Strain S7 was cultured in 5 ml of YM medium by shaking at 30.degree. C. After adding 0.55 ml of 10.times.TE (100 mM Tris-HCl, 10 mM EDTA, pH 8), 0.3 ml of 10% sodium dodecylsulfate, and 0.03 ml of 20 mg / ml proteinase K, the cultures were incubated with shaking for one hour at 65.degree. C. After adding 1 ml of 5M NaCi and 0.8 ml of 10% CTAB (hexadecyltrimethylammoniumbromide) in 1M NaCl the lysates were incubated 30 min at 65.degree. C., and then extracted once with chloroform and once with phenol:chloroform (1:1). The upper aqueous phase was removed and added to 0.6 volumes of isopropylalcohol and then dried. The precipitate was resuspended with a mixture of 0.6 ml of 1.times.TE containing 0.7 M NaCl and 0.1 ml of 10% CTAB in 1M NaCl, incubated 30 min at 65.degree. C., extracted once with chloroform, and then precipitated with two volumes of ethanol. After drying the pellet was resuspended in 0.1 ml of 1.times.TE. High mole...

example 4 (

NEW COMPOSITION)

As shown in Table 1, the broth viscosity for strain S7 carrying additional copies of the plasmid pRK-S7c6 was increased compared to that of unmodified strain S7. After purification of the exopolysaccharide from the broth, the EPS S7c6 retained its high viscosity as indicated in Table 2. EPS S7c6 is the exopolysaccharide produced by the Sphingomonas strain S7 carrying plasmid pRK-S7c6. The increased viscosity per gram of purified EPS suggested a new composition for the S7c6 polymer. The carbohydrate compositions for each of the exopolysaccharides from S7 and the genetically modified derivatives were determined following acid hydrolysis. The ratio of glucose to rhamnose is given in Table 2. The S7c6 EPS has a unique sugar composition with relatively less glucose residues. Table 2 shows that the new composition is linked to the high viscosity.

The carbohydrate compositions were determined for samples of the culture broths after precipitation of the EPS with 2 volumes of ...

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Abstract

A modified S7 polysaccharide is disclosed. The polysaccharide contains 20% less glucose that S7. An S7c6 gene cluster is disclosed. A mutated Sphingomonas is also disclosed.

Description

BACKGROUND LITERATUREPolysaccharide S-7 (abbreviated S-7) is the subject of four expired patents: (1) U.S. Pat. No. 3,960,832 issued to Kelco Jun. 1, 1976 and comprising a single composition of matter claim; (2) U.S. Pat. No. 3,915,800 issued to Kelco on Oct. 28, 1975 and comprising the growth of a naturally occurring bacterial strain Azotobacter indicus (deposited as ATCC 21423) in a submerged aerated culture in a nutrient medium and recovering the polysaccharide; (3) U.S. Pat. No. 3,894,976 issued on Jul. 15, 1975 and claiming use of S-7 in water based paints; and (4) U.S. Pat. No. 3,979,303 issued Sep. 7, 1976 and claiming use of S-7 in oil well drilling. A recent US patent (U.S. Pat. No. 5,772,912) issued Jun. 30, 1998 claims use of S-7 in anti-icing formulations, and another (U.S. Pat. No. 4,462,836) issued Jul. 31, 1984 claims use of S-7 in cement. The published literature concerning this polysaccharide is limited to a 1977 review by the Kelco inventors that is based on the in...

Claims

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Application Information

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IPC IPC(8): C08B37/00C12P19/04C12P19/00C12N1/20
CPCC08B37/006C12N1/20C12P19/04
Inventor POLLOCK, THOMAS J.
Owner SHIN ETSU BIO INC
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